Lipids and FA analysis of canine prostate tissue

It is widely reported that an association exists between dietary fat intake and the incidence of prostate cancer in humans. To study this association, there is a need for an animal model where prostate carcinogenesis occurs spontaneously. The canine prostate is considered a suitable experimental model for prostate cancer in humans since it is morphologically similar to the human prostate and both humans and dogs have a predisposition to benign and malignant prostate disease. In this study, the FA and lipids profiles of the normal canine prostate tissue from nine dogs were examined. The total lipid content of the canine prostate tissue was 1.7±0.5% (wet weight). The lipid composition analysis using TLC-FID showed that the two major lipid classes were phospholipids and TAG. Total FA, phospholipid, and TAG FA analysis showed that the major FA were palmitic acid (16∶0), stearic acid (18∶0), oleic acid (18∶1), linoleic acid (18∶2n−6), and arachidonic acid (20∶4n−6), The n−3 FA were present at <3% of total FA and included α-linolenic acid (18∶3n−3) (in total and TAG tissue FA), EPA (20∶5n−3) (not in TAG), and DHA (22∶6n−3) (not in TAG). The n−3/n−6 ratio was 1∶11, 1∶13, and 1∶8 in total, phospholipid, and TAG FA, respectively. This study shows the canine prostate has a low level of n−3 FA and a low n−3/n−6 ratio. This is perhaps due to low n−3 content of the diet of the dogs. FA analysis of dogfoods available in Australia showed that the n−3 content in both supermarket and premium bran dogfoods was <3% (wet weight), and the n−3/n−6 ratio was low.

Wireless estimation of canine pose for search and rescue

In this paper we discuss the use of accelerometers and Bluetooth to monitor canine pose in the context of common poses observed in urban search and rescue dogs. We discuss the use of the canine pose system in a disaster environment, and propose techniques for determining canine pose. In addition we discuss the challenges with this approach in such environments. The paper presents the experimental results obtained from the heavy urban search and rescue disaster simulation, where experiments were conducted using multiple canines, which show that angles can be derived from acceleration readings. Our experiments show that similar angles were measured for each of the poses, even when measured on multiple USAR canines of varying size. We also found measurable and consistent differences between each of the poses, making them clearly distinguishable from one another, again even when comparing with different USAR canines.

Mitomycin C: a promising agent for the treatment of canine corneal scarring

Objective  To evaluate the safety and efficacy of mitomycin C (MMC) in prevention of canine corneal scarring.

Methods  With an in vitro approach using healthy canine corneas, cultures of primary canine corneal fibroblasts or myofibroblasts were generated. Primary canine corneal fibroblasts were obtained by growing corneal buttons in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum-free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue assay and phase-contrast microscopy were used to evaluate the toxicity of three doses of MMC (0.002%, 0.02% and 0.04%). Real-time PCR, immunoblot, and immunocytochemistry techniques were used to determine MMC efficacy to inhibit markers of canine corneal scarring.

Results  A single 2-min treatment of 0.02% or less MMC did not alter canine corneal fibroblast or keratocyte phenotype, viability, or growth. The 0.02% dose substantially reduced myofibroblast formation (up to 67%; P < 0.001), as measured by the change in RNA and protein expression of fibrosis biomarkers (α-smooth muscle actin and F-actin).

Conclusion  This in vitro study suggests that a single 2-min 0.02% MMC treatment to the canine corneal keratocytes is safe and may be useful in decreasing canine corneal fibrous metaplasia. In vivo studies are warranted.

Characterization of haemolytic phospholipase A2 activity in clinical isolates of Campylobacter concisus

A membrane-bound, haemolytic phospholipase A2 (PLA2) activity was detected in clinical strains of Campylobacter concisus isolated from children with gastroenteritis. The clinical strains were assigned into two molecular groups (genomospecies) based on PCR amplification of their 23S rDNA. This calcium-dependent, heat-stable, haemolytic PLA2 activity was detected in strains from both genomospecies. A crude haemolysin extract (CHE) was initially prepared from cellular outer-membrane proteins of these isolates and was further fractionated by ultrafiltration. The haemolytic activity of the extracted fraction (R30) was retained by ultrafiltration using a 30 kDa molecular mass cut-off filter, and was designated haemolysin extract (HE). Both CHE and HE had PLA2 activity and caused stable vacuolating and cytolytic effects on Chinese hamster ovary cells in tissue culture. Primers for the conserved region of pldA gene (phospholipase A gene) from Campylobacter coli amplified a gene region of 460 bp in all tested isolates, confirming the presence of a homologous PLA gene sequence in C. concisus. The detection of haemolytic PLA₂ activity in C. concisus indicates the presence of a potential virulence factor in this species and supports the hypothesis that C. concisus is a possible opportunistic pathogen.

Domestic drinking water in rural areas : are water tanks on farms a health hazard?

The aim of the research was to gain a better understanding of the relationship between drinking water quality, householders' knowledge and maintenance practices of private water supplies and drinking water-related public health risk on farms. Samples of drinking water were taken from 100 farming households. The Colilert-18 method was used for the detection of total coliforms and Escherichia coli (E. coli) as indicators of water quality. Each household completed a questionnaire about their knowledge and practices relating to a safe water supply. Coliforms were present in 52 water samples and E. coli was present in 38. Seven households reported minor illnesses in the previous three months and two households reported gastroenteritis. Some tank maintenance occurred in 86 households, but tank maintenance activities varied considerably. Four of the households had published guidelines on water quality. None of the participating households had their drinking water tested regularly. There was no obvious relationship between drinking water quality, householder knowledge, maintenance practices and drinking water-related health risk on farms.

Characterization of Campylobacter concisus hemolysins

Campylobacter concisus is an opportunistic pathogen commonly found in the human oral cavity. It has also been isolated from clinical sources including gastroenteritis cases. Both secreted and cell-associated hemolytic activities were detected in C. concisus strains isolated from children with gastroenteritis. The secreted hemolytic activity of C. concisus strains was labile and was detected in variable levels from fresh-culture filtrates only. In addition, another secreted hemolysin/cytotoxin with a molecular weight <10 kDa was detected in a single C. concisus strain (RCH 12). A C. concisus genomic library, constructed from strain RCH 3 in Escherichia coli XL1-Blue, was screened for hemolytic clones. Subcloning and sequence analysis of selected hemolytic clones identified ORFs for genes that enhance hemolytic activity but do not appear to be related to any known hemolysin genes found in Gram-negative bacteria. In a previous study, a stable cell-associated hemolysin was identified as an outer-membrane phospholipase A (OMPLA) encoded by the pldA gene. In this study, we report cloning of the pldA gene of the clinical strain C. concisus RCH 3 and the complementation of phospholipase A activity in an E. coli pldA mutant.

Bone formation following implantation of titanium sponge rods into humeral osteotomies in dogs : a histological and histometrical study

Background: Titanium (Ti) is widely proven to enhance bone contact and growth on its surface. It is expected that bone defects could benefit from Ti to promote healing and to increase strength of the implanted area.

Purpose: The present study aimed at comparing the potential of porous Ti sponge rods with synthetic hydroxyapatite (HA) for the healing of bone defects in a canine model.

Material and Methods: Six mongrel dogs were submitted to three trephined osteotomies of 6.0 × 4.0 mm in one humerus and after 2 months another three osteotomies were performed in the contralateral humerus. A total of 36 defects were randomly filled either with Ti foam, particulate HA, or coagulum (control). The six animals were killed 4 months after the first surgery for histological and histometrical analysis.

Results: The Ti-foam surface was frequently found in intimate contact with new bone especially at the defect walls. Control sites showed higher amounts of newly formed bone at 2 months – Ti (p = 0.000) and HA (p = 0.009) – and 4 months when compared with Ti (p = 0.001). Differently from HA, the Ti foam was densely distributed across the defect area which rendered less space for bone growth in the latter's sites. The use of Ti foams or HA resulted in similar amounts of bone formation in both time intervals. Nevertheless, the presence of a Ti-foam rod preserved defect's marginal bone height as compared with control groups. Also, the Ti-foam group showed a more mature bone pattern at 4 months than HA sites.

Conclusion: The Ti foam exhibited good biocompatibility, and its application resulted in improved maintenance of bone height compared with control sites. The Ti foam in a rod design exhibited bone ingrowth properties suitable for further exploration in other experimental situations.

Diet and habitat of the powerful owl (Ninox strenua) living near Melbourne

The diet of Powerful Owls (Ninox strenua) living at Christmas Hills, 35km north-east of Melbourne was examined by analysis of 686 regurgitated pellets collected over two years. An aid was also developed to help identify potential mammalian prey species based on hair and skeletal characteristics. The following features were found to be most useful in distinguishing between the three species of arboreal marsupials - Common Ringtail Possum (Pseudocheirus peregrinus), Common Brushtail Possum (Trichosurus vulpecula) and Sugar Glider (Petaurus breviceps): - Cross-sectional width of primary guard hairs. - The size and shape of the nasal, frontal, parietal and squamosal bones of the skull. - Dentition. The size and shape of the upper incisor, canine and premolar teeth. The size and shape of the lower incisor and premolar teeth. - The size of the humerus. The Sugar Glider has a much smaller humerus than that of the Common Ringtail Possum and the Common Brushtail Possum. In the Common Brushtail Possum the entepicondyle ends in a very sharp point but the Common Ringtail Possum this point is not as sharp. - The Common Ringtail Possum’s femur has a very prominent trochanter which projects further than that in the Common Brushtail Possum. The femur of the Sugar Glider is distinguished by having a very large depression between the condyle and the trochanter. - The Common Brushtail Possum’s scapula has a narrower lower blade (relative to length) than that in the Common Ringtail Possum. The scapula of the Sugar Glider is smaller in size than that of the other two possums.The pelvic girdle Of the Common Brushtail Possum has a much wider ischium than those of the Common Brushtail Possum and the Sugar Glider. The ilium of the Sugar is much narrower and smaller than that of the other two possums Mammalian prey was found in 89%, insects in 13% and birds in 10% of the pellets. Of the mammals, Common Ringtail Possums occurred most frequently in the pellets over the year. There was no seasonal difference in the frequency of occurrence of Common Ringtail Possums and Sugar Gliders in pellets. However, Common Brushtail Possums were more likely to be taken in spring than in the other seasons. More adult Common Ringtail Possums were taken as prey than were other age classes over the year, except in summer when high numbers of young were consumed by the owls. The habitat of the Powerful Owl was examined by ground surveys and spotlight surveys in sixteen sites within the Warrandyte-Kinglake Nature Conservation Link. Four categories of survey sites were chosen with the following features. Category A - Sites with a dense understorey of shrubs and small trees, as well as many old trees (>10/ha) which might be suitable for nest hollows. Category B - Sites which lacked a dense understorey of shrubs and small trees and containing few or no old trees suitable for nest hollows. Category C - Sites with a dense understorey of shrubs and small trees but containing few or no old trees suitable for nest hollows. Category D - Sites which lacked a dense understorey of shrubs and small trees but having old trees (>10/ha) which might be suitable for nest hollows. High prey densities strongly correlated with the presence of hollows at these sites. In the light of the results, management recommendations were made for the future conservation of the Powerful Owls living at Christmas Hills. The following recommendations were particularly important: 1. Cleared or semi - cleared land within the Warrandyte Kinglake Nature Conservation Link be revegetated using indigenous species of eucalypts and waffles in order to provide a contiguous native forest corridor for the movement of possums and gliders between the Yarra River Valley and the Kinglake Plateau. 2. Continued planting of Eucalyptus spp. and Acacia spp. in the forested areas of the Warrandyte-Kinglake Nature Conservation Link. 3. Continued protection of healthy living trees to provide a continuous supply of hollow trees. 4. No falling of dead standing trees for firewood collecting as these can provide nest hollows for prey species of the Powerful Owl.

Relationship between alcohol abuse upper gastro-intestinal haemorrhage and nutritional status during acute hospital admission

This thesis is concerned with the effect of alcohol consumption on the pathogenesis of bleeding from the upper gastrointestinal tract via nutritional pathways. Altered nutritional status is a frequently recognised clinical accompaniement of heavy alcohol consumption in hospitalized patients. Similarly, upper gastrointestinal bleeding is frequently accompanied by the presence of heavy alcohol consumption. Nevertheless, the clinical quantification of alcohol intake is often descriptive, so that a link between alcohol use and upper gastrointestinal haemorrhage via nutritional mechanisms has been only generally defined. In the literature review, the methods of defining alcohol use and abuse, using interview, biochemical and haematological techniques are noted. The relationship between alcohol abuse and nutrient imbalances is reviewed, especially in relation to possible effects on the gastrointestinal tract, appetite and eating habits. A further section reviews the relationship between alcohol use and anatomical lesions of the upper gastrointestinal tract likely to lead to bleeding. Following the chapter in which the methods used in this thesis are described. Chapter 4 seeks to describe the study population and its subgroups in this thesis in relation to interview, biochemical and haematological methods. Alcohol use is defined in relation to (1) a clinical classification of heavy or light drinking, based on a questionnaire administered in Casualty, (2) a quantified method of determining alcohol consumption during a subsequent ward dietetic assessment, (3) in relation to a biochemical definition (recent drinking and non-drinking), and a classification of (1) and (2) called, for the purposes of this thesis, 'alcohol abusers' and 'nonabusers'. Heavy, regular and recent drinkers and alcohol abusers tend to be male and younger than light, infrequent and nonrecent drinkers and nonabusers. Chapter 5 relates the nutritional status of those patients admitted acutely to hospital in relation to the groups defined in Chapter 4, Nutritional status is defined in terms of food intake, anthropometry, biochemical and haematological parameters. Different methods of defining alcohol use give rise to different patterns of nutritional impairment. Chapter 6 relates the nutritional status of those patients admitted acutely to hospital in relation to the presence or absence of an endoscopically defined site of upper gastrointestinal bleeding. A difference is seen between those bleeding from a Mailory-weiss tear and other sites of bleeding, similarly, biochemical differences in nutritional status emerge between those patients who presented in shock, and those who did not. Chapter 7 explores the relationships between biochemical markers of nutritional status and haemostatic variables in the groups of abusers/non-abusers, the various sites of primary bleeding/controls, and shock/non-shock. Serum copper appears to be related to altered haemostasis in a manner not apparently described elsewhere.

Recent advances of metal binding protein lactoferrin as an anti-microbial agent

Lactoferrin (Lf) is present in milk and gland secretions and serve as an antimicrobial function. Insufficient amounts of Lf in some secretions also appear to correlate with certain health problems. Protection against gastroenteritis is the most likely biologically relevant activity of lactoferrin. Multiple in vitro and animal studies have shown a protective effect of lactoferrin on infections with enteric microorganisms, including rotavirus, Giardia, Shigella, Salmonella and the diarrheagenic Escherichia coli. Lactoferrin has two major effects on enteric pathogens: it inhibits growth and it impairs function of surface expressed virulence factors thereby decreasing their ability to adhere or to invade mammalian cells. Lf also inhibits several species of fungi and certain parasites. This review covers the role of Lf in clearing the parasitic infections. The mechanism by which lactoferrin inhibits some parasites may be via stimulation of the process of phagocytosis, whereby immune cells engulf and digest foreign organisms. Trichomonas vaginalis is a protozoan responsible for the number one, non-viral sexually transmitted disease. In this review, we also discussed the role of Lf in cervical infections.

Chemotoxicity of doxorubicin and surface expression of P-glycoprotein (MDR1) is regulated by the Pseudomonas aeruginosa toxin Cif

P-glycoprotein (Pgp), a member of the adenosine triphosphate-binding cassette (ABC) transporter superfamily, is a major drug efflux pump expressed in normal tissues, and is overexpressed in many human cancers. Overexpression of Pgp results in reduced intracellular drug concentration and cytotoxicity of chemotherapeutic drugs and is thought to contribute to multidrug resistance of cancer cells. The involvement of Pgp in clinical drug resistance has led to a search for molecules that block Pgp transporter activity to improve the efficacy and pharmacokinetics of therapeutic agents. We have recently identified and characterized a secreted toxin from Pseudomonas aeruginosa, designated cystic fibrosis transmembrane conductance regulator (CFTR) inhibitory factor (Cif). Cif reduces the apical membrane abundance of CFTR, also an ABC transporter, and inhibits the CFTR-mediated chloride ion secretion by human airway and kidney epithelial cells. We report presently that Cif also inhibits the apical membrane abundance of Pgp in kidney, airway, and intestinal epithelial cells but has no effect on plasma membrane abundance of multidrug resistance protein 1 or 2. Cif increased the drug sensitivity to doxorubicin in kidney cells expressing Pgp by 10-fold and increased the cellular accumulation of daunorubicin by 2-fold. Thus our studies show that Cif increases the sensitivity of Pgp-overexpressing cells to doxorubicin, consistent with the hypothesis that Cif affects Pgp functional expression. These results suggest that Cif may be useful to develop a new class of specific inhibitors of Pgp aimed at increasing the sensitivity of tumors to chemotherapeutic drugs, and at improving the bioavailability of Pgp transport substrates.

Shedding new light on old species identifications : morphological and genetic evidence suggest a need for conservation status review of the critically endangered bat, Saccolaimus saccolaimus

Information based on the accurate identification of species is a vital component for achieving successful outcomes of biodiversity conservation and management. It is difficult to manage species that are poorly known or that are misidentified with other similar species. This is particularly problematic for rare and threatened species. Species that are listed under endangered species classification schemes need to be identified accurately and categorised correctly so that conservation efforts are appropriately allocated. In Australia, the emballonurid Saccolaimus saccolaimus is currently listed as ‘Critically Endangered’. On the basis of new observations and existing museum specimens, we used a combination of genetic (mitochondrial DNA sequence) and morphological (pelage characteristics, dig III : phalanx I length ratio, inter-upper canine distance) analyses to identify six new geographic records for S. saccolaimus, comprising ~100 individuals. Our analyses also suggested that there are likely to be more records in museum collections misidentified as S. flaviventris specimens. The external morphological similarities to S. flaviventris were addressed and genetic, morphological and echolocation analyses were used in an attempt to provide diagnostic characters that can be used to readily identify the two species in the field. We recommend genetic testing of all museum specimens of Australian Saccolaimus to clarify species’ distributions and provide data for reassessing the conservation status for both S. saccolaimus and S. flaviventris. Museum curators, taxonomists and wildlife managers need to be aware of potential species misidentifications, both in the field and laboratory. Misidentifications that result in misclassification of both threatened and non-threatened species can have significant implications.